Identification and application of a novel strong constitutive promoter in Corynebacterium glutamicum
Abstract The replacement of promoters with various strengths is an effective strategy to fine-tune gene expression. More available promoters with a broad range of transcription efficiency are needed in metabolic engineering. In this study, a putative protein coding gene CP_2454 was identified with a...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Wei, Hongbo [verfasserIn] |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2018 |
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| Schlagwörter: |
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| Anmerkung: |
© Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018 |
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| Übergeordnetes Werk: |
Enthalten in: Annals of microbiology - Berlin : Springer, 1998, 68(2018), 6 vom: 11. Mai, Seite 375-382 |
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| Übergeordnetes Werk: |
volume:68 ; year:2018 ; number:6 ; day:11 ; month:05 ; pages:375-382 |
| Links: |
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| DOI / URN: |
10.1007/s13213-018-1344-0 |
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| Katalog-ID: |
SPR030889529 |
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| 245 | 1 | 0 | |a Identification and application of a novel strong constitutive promoter in Corynebacterium glutamicum |
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| 520 | |a Abstract The replacement of promoters with various strengths is an effective strategy to fine-tune gene expression. More available promoters with a broad range of transcription efficiency are needed in metabolic engineering. In this study, a putative protein coding gene CP_2454 was identified with a stable and high transcriptional level from an l-leucine-producing strain Corynebacterium glutamicum CP, which was absent in wild-type C. glutamicum ATCC 13032. The transcriptional level of CP_2454 was about 80.0% those of tuf and sod, and was 1.6 and 3.2 times those of ilvB and gapA, respectively. These promoters were cloned into C. glutamicum ATCC 13032 to control the expression of green fluorescent protein (GFP). While, the expressional level of GFP under the control of $ P_{CP_2454} $ was close to that of $ P_{tuf} $ and $ P_{sod} $, which was significantly higher than that of other tested promoters. The native promoter of ilvB controlling the expression of a feedback-resistant acetolactate synthase was replaced by $ P_{CP_2454} $, resulting in an increase of l-valine titer by 58.5%. A further 24.9% increase of l-valine titer was achieved after replacement of the promoter of ilvD encoding dihydroxyacid dehydratase with $ P_{CP_2454} $. Identification of the constitutive promoter $ P_{CP_2454} $ expands the promoter library of C. glutamicum and provides essential reference for tunable expression of target genes in C. glutamicum. | ||
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| 700 | 1 | |a Chen, Ning |4 aut | |
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10.1007/s13213-018-1344-0 doi (DE-627)SPR030889529 (SPR)s13213-018-1344-0-e DE-627 ger DE-627 rakwb eng Wei, Hongbo verfasserin aut Identification and application of a novel strong constitutive promoter in Corynebacterium glutamicum 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018 Abstract The replacement of promoters with various strengths is an effective strategy to fine-tune gene expression. More available promoters with a broad range of transcription efficiency are needed in metabolic engineering. In this study, a putative protein coding gene CP_2454 was identified with a stable and high transcriptional level from an l-leucine-producing strain Corynebacterium glutamicum CP, which was absent in wild-type C. glutamicum ATCC 13032. The transcriptional level of CP_2454 was about 80.0% those of tuf and sod, and was 1.6 and 3.2 times those of ilvB and gapA, respectively. These promoters were cloned into C. glutamicum ATCC 13032 to control the expression of green fluorescent protein (GFP). While, the expressional level of GFP under the control of $ P_{CP_2454} $ was close to that of $ P_{tuf} $ and $ P_{sod} $, which was significantly higher than that of other tested promoters. The native promoter of ilvB controlling the expression of a feedback-resistant acetolactate synthase was replaced by $ P_{CP_2454} $, resulting in an increase of l-valine titer by 58.5%. A further 24.9% increase of l-valine titer was achieved after replacement of the promoter of ilvD encoding dihydroxyacid dehydratase with $ P_{CP_2454} $. Identification of the constitutive promoter $ P_{CP_2454} $ expands the promoter library of C. glutamicum and provides essential reference for tunable expression of target genes in C. glutamicum. Constitutive promoter (dpeaa)DE-He213 Transcriptional level (dpeaa)DE-He213 Fluorescence intensity (dpeaa)DE-He213 Enzyme expression (dpeaa)DE-He213 -valine (dpeaa)DE-He213 Ma, Yuechao aut Chen, Qixin aut Cui, Yi aut Du, Lihong aut Ma, Qian aut Li, Yanjun aut Xie, Xixian aut Chen, Ning aut Enthalten in Annals of microbiology Berlin : Springer, 1998 68(2018), 6 vom: 11. Mai, Seite 375-382 (DE-627)385615434 (DE-600)2143009-3 1869-2044 nnns volume:68 year:2018 number:6 day:11 month:05 pages:375-382 https://dx.doi.org/10.1007/s13213-018-1344-0 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_22 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_63 GBV_ILN_95 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_187 GBV_ILN_285 GBV_ILN_370 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2007 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2070 GBV_ILN_2086 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2116 GBV_ILN_2118 GBV_ILN_2119 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4313 GBV_ILN_4328 GBV_ILN_4333 AR 68 2018 6 11 05 375-382 |
| spelling |
10.1007/s13213-018-1344-0 doi (DE-627)SPR030889529 (SPR)s13213-018-1344-0-e DE-627 ger DE-627 rakwb eng Wei, Hongbo verfasserin aut Identification and application of a novel strong constitutive promoter in Corynebacterium glutamicum 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018 Abstract The replacement of promoters with various strengths is an effective strategy to fine-tune gene expression. More available promoters with a broad range of transcription efficiency are needed in metabolic engineering. In this study, a putative protein coding gene CP_2454 was identified with a stable and high transcriptional level from an l-leucine-producing strain Corynebacterium glutamicum CP, which was absent in wild-type C. glutamicum ATCC 13032. The transcriptional level of CP_2454 was about 80.0% those of tuf and sod, and was 1.6 and 3.2 times those of ilvB and gapA, respectively. These promoters were cloned into C. glutamicum ATCC 13032 to control the expression of green fluorescent protein (GFP). While, the expressional level of GFP under the control of $ P_{CP_2454} $ was close to that of $ P_{tuf} $ and $ P_{sod} $, which was significantly higher than that of other tested promoters. The native promoter of ilvB controlling the expression of a feedback-resistant acetolactate synthase was replaced by $ P_{CP_2454} $, resulting in an increase of l-valine titer by 58.5%. A further 24.9% increase of l-valine titer was achieved after replacement of the promoter of ilvD encoding dihydroxyacid dehydratase with $ P_{CP_2454} $. Identification of the constitutive promoter $ P_{CP_2454} $ expands the promoter library of C. glutamicum and provides essential reference for tunable expression of target genes in C. glutamicum. Constitutive promoter (dpeaa)DE-He213 Transcriptional level (dpeaa)DE-He213 Fluorescence intensity (dpeaa)DE-He213 Enzyme expression (dpeaa)DE-He213 -valine (dpeaa)DE-He213 Ma, Yuechao aut Chen, Qixin aut Cui, Yi aut Du, Lihong aut Ma, Qian aut Li, Yanjun aut Xie, Xixian aut Chen, Ning aut Enthalten in Annals of microbiology Berlin : Springer, 1998 68(2018), 6 vom: 11. Mai, Seite 375-382 (DE-627)385615434 (DE-600)2143009-3 1869-2044 nnns volume:68 year:2018 number:6 day:11 month:05 pages:375-382 https://dx.doi.org/10.1007/s13213-018-1344-0 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_22 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_63 GBV_ILN_95 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_187 GBV_ILN_285 GBV_ILN_370 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2007 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2070 GBV_ILN_2086 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2116 GBV_ILN_2118 GBV_ILN_2119 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4313 GBV_ILN_4328 GBV_ILN_4333 AR 68 2018 6 11 05 375-382 |
| allfields_unstemmed |
10.1007/s13213-018-1344-0 doi (DE-627)SPR030889529 (SPR)s13213-018-1344-0-e DE-627 ger DE-627 rakwb eng Wei, Hongbo verfasserin aut Identification and application of a novel strong constitutive promoter in Corynebacterium glutamicum 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018 Abstract The replacement of promoters with various strengths is an effective strategy to fine-tune gene expression. More available promoters with a broad range of transcription efficiency are needed in metabolic engineering. In this study, a putative protein coding gene CP_2454 was identified with a stable and high transcriptional level from an l-leucine-producing strain Corynebacterium glutamicum CP, which was absent in wild-type C. glutamicum ATCC 13032. The transcriptional level of CP_2454 was about 80.0% those of tuf and sod, and was 1.6 and 3.2 times those of ilvB and gapA, respectively. These promoters were cloned into C. glutamicum ATCC 13032 to control the expression of green fluorescent protein (GFP). While, the expressional level of GFP under the control of $ P_{CP_2454} $ was close to that of $ P_{tuf} $ and $ P_{sod} $, which was significantly higher than that of other tested promoters. The native promoter of ilvB controlling the expression of a feedback-resistant acetolactate synthase was replaced by $ P_{CP_2454} $, resulting in an increase of l-valine titer by 58.5%. A further 24.9% increase of l-valine titer was achieved after replacement of the promoter of ilvD encoding dihydroxyacid dehydratase with $ P_{CP_2454} $. Identification of the constitutive promoter $ P_{CP_2454} $ expands the promoter library of C. glutamicum and provides essential reference for tunable expression of target genes in C. glutamicum. Constitutive promoter (dpeaa)DE-He213 Transcriptional level (dpeaa)DE-He213 Fluorescence intensity (dpeaa)DE-He213 Enzyme expression (dpeaa)DE-He213 -valine (dpeaa)DE-He213 Ma, Yuechao aut Chen, Qixin aut Cui, Yi aut Du, Lihong aut Ma, Qian aut Li, Yanjun aut Xie, Xixian aut Chen, Ning aut Enthalten in Annals of microbiology Berlin : Springer, 1998 68(2018), 6 vom: 11. Mai, Seite 375-382 (DE-627)385615434 (DE-600)2143009-3 1869-2044 nnns volume:68 year:2018 number:6 day:11 month:05 pages:375-382 https://dx.doi.org/10.1007/s13213-018-1344-0 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_22 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_63 GBV_ILN_95 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_187 GBV_ILN_285 GBV_ILN_370 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2007 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2070 GBV_ILN_2086 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2116 GBV_ILN_2118 GBV_ILN_2119 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4313 GBV_ILN_4328 GBV_ILN_4333 AR 68 2018 6 11 05 375-382 |
| allfieldsGer |
10.1007/s13213-018-1344-0 doi (DE-627)SPR030889529 (SPR)s13213-018-1344-0-e DE-627 ger DE-627 rakwb eng Wei, Hongbo verfasserin aut Identification and application of a novel strong constitutive promoter in Corynebacterium glutamicum 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018 Abstract The replacement of promoters with various strengths is an effective strategy to fine-tune gene expression. More available promoters with a broad range of transcription efficiency are needed in metabolic engineering. In this study, a putative protein coding gene CP_2454 was identified with a stable and high transcriptional level from an l-leucine-producing strain Corynebacterium glutamicum CP, which was absent in wild-type C. glutamicum ATCC 13032. The transcriptional level of CP_2454 was about 80.0% those of tuf and sod, and was 1.6 and 3.2 times those of ilvB and gapA, respectively. These promoters were cloned into C. glutamicum ATCC 13032 to control the expression of green fluorescent protein (GFP). While, the expressional level of GFP under the control of $ P_{CP_2454} $ was close to that of $ P_{tuf} $ and $ P_{sod} $, which was significantly higher than that of other tested promoters. The native promoter of ilvB controlling the expression of a feedback-resistant acetolactate synthase was replaced by $ P_{CP_2454} $, resulting in an increase of l-valine titer by 58.5%. A further 24.9% increase of l-valine titer was achieved after replacement of the promoter of ilvD encoding dihydroxyacid dehydratase with $ P_{CP_2454} $. Identification of the constitutive promoter $ P_{CP_2454} $ expands the promoter library of C. glutamicum and provides essential reference for tunable expression of target genes in C. glutamicum. Constitutive promoter (dpeaa)DE-He213 Transcriptional level (dpeaa)DE-He213 Fluorescence intensity (dpeaa)DE-He213 Enzyme expression (dpeaa)DE-He213 -valine (dpeaa)DE-He213 Ma, Yuechao aut Chen, Qixin aut Cui, Yi aut Du, Lihong aut Ma, Qian aut Li, Yanjun aut Xie, Xixian aut Chen, Ning aut Enthalten in Annals of microbiology Berlin : Springer, 1998 68(2018), 6 vom: 11. Mai, Seite 375-382 (DE-627)385615434 (DE-600)2143009-3 1869-2044 nnns volume:68 year:2018 number:6 day:11 month:05 pages:375-382 https://dx.doi.org/10.1007/s13213-018-1344-0 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_22 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_63 GBV_ILN_95 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_187 GBV_ILN_285 GBV_ILN_370 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2007 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2070 GBV_ILN_2086 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2116 GBV_ILN_2118 GBV_ILN_2119 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4313 GBV_ILN_4328 GBV_ILN_4333 AR 68 2018 6 11 05 375-382 |
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10.1007/s13213-018-1344-0 doi (DE-627)SPR030889529 (SPR)s13213-018-1344-0-e DE-627 ger DE-627 rakwb eng Wei, Hongbo verfasserin aut Identification and application of a novel strong constitutive promoter in Corynebacterium glutamicum 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018 Abstract The replacement of promoters with various strengths is an effective strategy to fine-tune gene expression. More available promoters with a broad range of transcription efficiency are needed in metabolic engineering. In this study, a putative protein coding gene CP_2454 was identified with a stable and high transcriptional level from an l-leucine-producing strain Corynebacterium glutamicum CP, which was absent in wild-type C. glutamicum ATCC 13032. The transcriptional level of CP_2454 was about 80.0% those of tuf and sod, and was 1.6 and 3.2 times those of ilvB and gapA, respectively. These promoters were cloned into C. glutamicum ATCC 13032 to control the expression of green fluorescent protein (GFP). While, the expressional level of GFP under the control of $ P_{CP_2454} $ was close to that of $ P_{tuf} $ and $ P_{sod} $, which was significantly higher than that of other tested promoters. The native promoter of ilvB controlling the expression of a feedback-resistant acetolactate synthase was replaced by $ P_{CP_2454} $, resulting in an increase of l-valine titer by 58.5%. A further 24.9% increase of l-valine titer was achieved after replacement of the promoter of ilvD encoding dihydroxyacid dehydratase with $ P_{CP_2454} $. Identification of the constitutive promoter $ P_{CP_2454} $ expands the promoter library of C. glutamicum and provides essential reference for tunable expression of target genes in C. glutamicum. Constitutive promoter (dpeaa)DE-He213 Transcriptional level (dpeaa)DE-He213 Fluorescence intensity (dpeaa)DE-He213 Enzyme expression (dpeaa)DE-He213 -valine (dpeaa)DE-He213 Ma, Yuechao aut Chen, Qixin aut Cui, Yi aut Du, Lihong aut Ma, Qian aut Li, Yanjun aut Xie, Xixian aut Chen, Ning aut Enthalten in Annals of microbiology Berlin : Springer, 1998 68(2018), 6 vom: 11. Mai, Seite 375-382 (DE-627)385615434 (DE-600)2143009-3 1869-2044 nnns volume:68 year:2018 number:6 day:11 month:05 pages:375-382 https://dx.doi.org/10.1007/s13213-018-1344-0 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_22 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_63 GBV_ILN_95 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_152 GBV_ILN_187 GBV_ILN_285 GBV_ILN_370 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2007 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2055 GBV_ILN_2059 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2070 GBV_ILN_2086 GBV_ILN_2106 GBV_ILN_2108 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2116 GBV_ILN_2118 GBV_ILN_2119 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2190 GBV_ILN_2232 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4313 GBV_ILN_4328 GBV_ILN_4333 AR 68 2018 6 11 05 375-382 |
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Wei, Hongbo Ma, Yuechao Chen, Qixin Cui, Yi Du, Lihong Ma, Qian Li, Yanjun Xie, Xixian Chen, Ning |
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Elektronische Aufsätze |
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Wei, Hongbo |
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10.1007/s13213-018-1344-0 |
| title_sort |
identification and application of a novel strong constitutive promoter in corynebacterium glutamicum |
| title_auth |
Identification and application of a novel strong constitutive promoter in Corynebacterium glutamicum |
| abstract |
Abstract The replacement of promoters with various strengths is an effective strategy to fine-tune gene expression. More available promoters with a broad range of transcription efficiency are needed in metabolic engineering. In this study, a putative protein coding gene CP_2454 was identified with a stable and high transcriptional level from an l-leucine-producing strain Corynebacterium glutamicum CP, which was absent in wild-type C. glutamicum ATCC 13032. The transcriptional level of CP_2454 was about 80.0% those of tuf and sod, and was 1.6 and 3.2 times those of ilvB and gapA, respectively. These promoters were cloned into C. glutamicum ATCC 13032 to control the expression of green fluorescent protein (GFP). While, the expressional level of GFP under the control of $ P_{CP_2454} $ was close to that of $ P_{tuf} $ and $ P_{sod} $, which was significantly higher than that of other tested promoters. The native promoter of ilvB controlling the expression of a feedback-resistant acetolactate synthase was replaced by $ P_{CP_2454} $, resulting in an increase of l-valine titer by 58.5%. A further 24.9% increase of l-valine titer was achieved after replacement of the promoter of ilvD encoding dihydroxyacid dehydratase with $ P_{CP_2454} $. Identification of the constitutive promoter $ P_{CP_2454} $ expands the promoter library of C. glutamicum and provides essential reference for tunable expression of target genes in C. glutamicum. © Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018 |
| abstractGer |
Abstract The replacement of promoters with various strengths is an effective strategy to fine-tune gene expression. More available promoters with a broad range of transcription efficiency are needed in metabolic engineering. In this study, a putative protein coding gene CP_2454 was identified with a stable and high transcriptional level from an l-leucine-producing strain Corynebacterium glutamicum CP, which was absent in wild-type C. glutamicum ATCC 13032. The transcriptional level of CP_2454 was about 80.0% those of tuf and sod, and was 1.6 and 3.2 times those of ilvB and gapA, respectively. These promoters were cloned into C. glutamicum ATCC 13032 to control the expression of green fluorescent protein (GFP). While, the expressional level of GFP under the control of $ P_{CP_2454} $ was close to that of $ P_{tuf} $ and $ P_{sod} $, which was significantly higher than that of other tested promoters. The native promoter of ilvB controlling the expression of a feedback-resistant acetolactate synthase was replaced by $ P_{CP_2454} $, resulting in an increase of l-valine titer by 58.5%. A further 24.9% increase of l-valine titer was achieved after replacement of the promoter of ilvD encoding dihydroxyacid dehydratase with $ P_{CP_2454} $. Identification of the constitutive promoter $ P_{CP_2454} $ expands the promoter library of C. glutamicum and provides essential reference for tunable expression of target genes in C. glutamicum. © Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018 |
| abstract_unstemmed |
Abstract The replacement of promoters with various strengths is an effective strategy to fine-tune gene expression. More available promoters with a broad range of transcription efficiency are needed in metabolic engineering. In this study, a putative protein coding gene CP_2454 was identified with a stable and high transcriptional level from an l-leucine-producing strain Corynebacterium glutamicum CP, which was absent in wild-type C. glutamicum ATCC 13032. The transcriptional level of CP_2454 was about 80.0% those of tuf and sod, and was 1.6 and 3.2 times those of ilvB and gapA, respectively. These promoters were cloned into C. glutamicum ATCC 13032 to control the expression of green fluorescent protein (GFP). While, the expressional level of GFP under the control of $ P_{CP_2454} $ was close to that of $ P_{tuf} $ and $ P_{sod} $, which was significantly higher than that of other tested promoters. The native promoter of ilvB controlling the expression of a feedback-resistant acetolactate synthase was replaced by $ P_{CP_2454} $, resulting in an increase of l-valine titer by 58.5%. A further 24.9% increase of l-valine titer was achieved after replacement of the promoter of ilvD encoding dihydroxyacid dehydratase with $ P_{CP_2454} $. Identification of the constitutive promoter $ P_{CP_2454} $ expands the promoter library of C. glutamicum and provides essential reference for tunable expression of target genes in C. glutamicum. © Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018 |
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6 |
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Identification and application of a novel strong constitutive promoter in Corynebacterium glutamicum |
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https://dx.doi.org/10.1007/s13213-018-1344-0 |
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Ma, Yuechao Chen, Qixin Cui, Yi Du, Lihong Ma, Qian Li, Yanjun Xie, Xixian Chen, Ning |
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